Journal of Lipid Research, Vol 33, 273-280, Copyright © 1992 by Lipid Research, Inc.
Immunological discrimination between the human apolipoprotein E2(Arg158- ---Cys) and E3 isoforms
K Gerritse, P de Knijff, G van Ierssel, LM Havekes, RR Frants, MM Schellekens, ND Zegers, E Claassen and WJ Boersma
Medical Biological Laboratory TNO, Rijswijk, The Netherlands.
A specific anti-apoE2(Arg158----Cys) monoclonal antibody was raised by
means of immunization of mice with a variant specific synthetic peptide.
The peptide sequences used were homologous to apolipoprotein E of human and
mouse. Consequently, the mouse immune system was tolerant to most of the
selected sequences. Immunization with only one of selected peptides (amino
acids 154-172) evoked an anti-peptide and anti- native protein response.
Surprisingly, this peptide was predicted to have a low antigenicity index,
in contrast to the other used peptides. The variant specific anti-peptide
MAb that was generated with this sequence, recognizes apoE2(Arg158----Cys)
and not apoE3. We here describe a sensitive, time saving, and simple
immunoblot assay to detect apoE2(Arg158----Cys) in human sera without prior
isoelectric focusing of serum proteins.
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